Universal solid supports for the synthesis of oligonucleotides with terminal 3′-phosphates

Abstract

The preparation of two types of supports based on controlled pore glass (CPG) is presented. These supports are compatible with established phosphoroamidite chemistry of oligonucleotides synthesis giving rise to an oligonucleotide with terminal 3'-phosphate function during final deprotection. CPG was modified with: (i) methacrylic acid derivatives and 2-mercaptoethanol (1) or (ii) aminoalkylsilane, succinic anhydride and benzidine (2). Support 2 can be also used for the synthesis of partially protected oligonucleotide 3'-phosphates.

Polymer supported synthesis of aminooxyalkylated oligonucleotides, and some applications in the fabrication of microarrays

“…In order to study the effect of pH on immobilization of aminooxyalkylated oligomers on epoxylated glass slides, the attachment of probe was carried out at different pH (4)(5)(6)(7)(8)(9)(10)(11). A labeled oligomer, TET-d(TTC GTC ATC AAG TAG TTC CT)-OPO 3 -CH 2 CH(OH)-CH 2 NHCOCH 2 ONH 2, was dissolved in reaction buffers having different pH and spotted (0.5 ll) onto epoxylated glass slides at a concentration of 1 lM using a pipettmen followed by overnight incubation at 45°C.…”

“…The technology allows highly parallel throughput processing and screening of different biological samples (e.g., mRNA, cDNA and proteins). [7][8][9][10][11][12][13][14][15] DNA microarrays are mostly based on hybridization of targets with double-stranded DNA or single-stranded oligonucleotides (probes) immobilized on a polymer support (usually glass), followed by detection by a scanning process. Of the two established methods, viz., on-chip synthesis and deposition method, the latter is the preferred route for the preparation of oligonucleotide microarrays for routine applications, as it offers flexibility in respect of different chemistries for attachment of probes on a surface of choice.…”

Solid-phase synthesis of base-sensitive oligonucleotides

“…Phosphoramidate linkage 24 deserves special attention, since its simplicity. It may be cleaved by fluoride ion (Et 3 N•3HF-THF, 1:1, v/v, 4h at rt) acting like a silyl linker (11)(12)(13)(14), 65 …”

Solid‐Phase Supports for Oligonucleotide Synthesis

“…3.1.4C and K). (A) Benzidine linker arm (Markiewicz and Wyrzykiewicz, 1989). (B) Hydroxyethylsulfonyl linker arm (Markiewicz and Wyrzykiewicz, 1989).…”